A Reportagem na tv: Caco Barcellos: um repórter e a injustiça social

Destacar a importância da reportagem investigativa, e mostrar o valor que deve ser observado no empenho da atuação profissional ética de um repórter de televisão, capaz de mudar o rumo da história política recente do Brasil, é o que se propõe o presente trabalho. A prática do jornalismo autêntico demonstra ser transformadora. É o que acontece com a atuação do repórter Caco Barcellos. Ele se propôs a perseguir uma história por mais de um ano e, ao final, mostrar uma verdade escondida durante 30 anos pelo poder público. Com tal reportagem, possibilitou a reparação de uma injustiça social cometida contra uma jovem estudante brasileira. A força da reportagem apresentada num telejornal é incontestável, em razão do potencial que representa a televisão na penetração em todas as camadas da sociedade como veículo de informação. Sendo a reportagem apresentada em série de três dias consecutivos pelo Jornal Nacional, eleva seu peso significativamente, devido à liderança de audiência do telejornal da Rede Globo. O investimento da emissora numa reportagem como a apresentada neste trabalho depende fundamentalmente da presença de um repórter profundamente identificado com a luta contra a injustiça social. E é este o perfil de Caco Barcellos. Ele cresceu na periferia pobre de uma metrópole, e desde muito cedo se revoltou contra a ação policial que discrimina o cidadão comum e privilegia os poderosos. Conseguiu transformar a raiva acumulada em ações efetivas através da reportagem investigativa reveladora da injustiça social cometida diariamente. Quando o espaço era pequeno para o tamanho do que tinha para reportar, Caco lançou livros, ganhou prêmios e enfrentou processos judiciais, movidos por policiais interessados em destruir a vida profissional do repórter que ousou trabalhar sério e encontrar os documentos que provavam os desmandos praticados contra inocentes, friamente assassinados. O resultado é uma atuação exemplar, com um método de trabalho digno de ser seguido, refletindo-se na série de reportagens que desmontou uma farsa montada pelo exército, e que pode ser considerada como uma verdadeira aula de jornalismo.

A Reportagem na tv: Caco Barcellos: um repórter e a injustiça social

Destacar a importância da reportagem investigativa, e mostrar o valor que deve ser observado no empenho da atuação profissional ética de um repórter de televisão, capaz de mudar o rumo da história política recente do Brasil, é o que se propõe o presente trabalho. A prática do jornalismo autêntico demonstra ser transformadora. É o que acontece com a atuação do repórter Caco Barcellos. Ele se propôs a perseguir uma história por mais de um ano e, ao final, mostrar uma verdade escondida durante 30 anos pelo poder público. Com tal reportagem, possibilitou a reparação de uma injustiça social cometida contra uma jovem estudante brasileira. A força da reportagem apresentada num telejornal é incontestável, em razão do potencial que representa a televisão na penetração em todas as camadas da sociedade como veículo de informação. Sendo a reportagem apresentada em série de três dias consecutivos pelo Jornal Nacional, eleva seu peso significativamente, devido à liderança de audiência do telejornal da Rede Globo. O investimento da emissora numa reportagem como a apresentada neste trabalho depende fundamentalmente da presença de um repórter profundamente identificado com a luta contra a injustiça social. E é este o perfil de Caco Barcellos. Ele cresceu na periferia pobre de uma metrópole, e desde muito cedo se revoltou contra a ação policial que discrimina o cidadão comum e privilegia os poderosos. Conseguiu transformar a raiva acumulada em ações efetivas através da reportagem investigativa reveladora da injustiça social cometida diariamente. Quando o espaço era pequeno para o tamanho do que tinha para reportar, Caco lançou livros, ganhou prêmios e enfrentou processos judiciais, movidos por policiais interessados em destruir a vida profissional do repórter que ousou trabalhar sério e encontrar os documentos que provavam os desmandos praticados contra inocentes, friamente assassinados. O resultado é uma atuação exemplar, com um método de trabalho digno de ser seguido, refletindo-se na série de reportagens que desmontou uma farsa montada pelo exército, e que pode ser considerada como uma verdadeira aula de jornalismo.

A Reportagem na tv: Caco Barcellos: um repórter e a injustiça social

Destacar a importância da reportagem investigativa, e mostrar o valor que deve ser observado no empenho da atuação profissional ética de um repórter de televisão, capaz de mudar o rumo da história política recente do Brasil, é o que se propõe o presente trabalho. A prática do jornalismo autêntico demonstra ser transformadora. É o que acontece com a atuação do repórter Caco Barcellos. Ele se propôs a perseguir uma história por mais de um ano e, ao final, mostrar uma verdade escondida durante 30 anos pelo poder público. Com tal reportagem, possibilitou a reparação de uma injustiça social cometida contra uma jovem estudante brasileira. A força da reportagem apresentada num telejornal é incontestável, em razão do potencial que representa a televisão na penetração em todas as camadas da sociedade como veículo de informação. Sendo a reportagem apresentada em série de três dias consecutivos pelo Jornal Nacional, eleva seu peso significativamente, devido à liderança de audiência do telejornal da Rede Globo. O investimento da emissora numa reportagem como a apresentada neste trabalho depende fundamentalmente da presença de um repórter profundamente identificado com a luta contra a injustiça social. E é este o perfil de Caco Barcellos. Ele cresceu na periferia pobre de uma metrópole, e desde muito cedo se revoltou contra a ação policial que discrimina o cidadão comum e privilegia os poderosos. Conseguiu transformar a raiva acumulada em ações efetivas através da reportagem investigativa reveladora da injustiça social cometida diariamente. Quando o espaço era pequeno para o tamanho do que tinha para reportar, Caco lançou livros, ganhou prêmios e enfrentou processos judiciais, movidos por policiais interessados em destruir a vida profissional do repórter que ousou trabalhar sério e encontrar os documentos que provavam os desmandos praticados contra inocentes, friamente assassinados. O resultado é uma atuação exemplar, com um método de trabalho digno de ser seguido, refletindo-se na série de reportagens que desmontou uma farsa montada pelo exército, e que pode ser considerada como uma verdadeira aula de jornalismo.

Solid Lipid Nanoparticles For Hydrophilic Biotech Drugs: Optimization And Cell Viability Studies (caco-2 & Hepg-2 Cell Lines).

Insulin was used as model protein to developed innovative Solid Lipid Nanoparticles (SLNs) for the delivery of hydrophilic biotech drugs, with potential use in medicinal chemistry. SLNs were prepared by double emulsion with the purpose of promoting stability and enhancing the protein bioavailability. Softisan(®)100 was selected as solid lipid matrix. The surfactants (Tween(®)80, Span(®)80 and Lipoid(®)S75) and insulin were chosen applying a 2(2) factorial design with triplicate of central point, evaluating the influence of dependents variables as polydispersity index (PI), mean particle size (z-AVE), zeta potential (ZP) and encapsulation efficiency (EE) by factorial design using the ANOVA test. Therefore, thermodynamic stability, polymorphism and matrix crystallinity were checked by Differential Scanning Calorimetry (DSC) and Wide Angle X-ray Diffraction (WAXD), whereas the effect of toxicity of SLNs was check in HepG2 and Caco-2 cells. Results showed a mean particle size (z-AVE) width between 294.6 nm and 627.0 nm, a PI in the range of 0.425-0.750, ZP about -3 mV, and the EE between 38.39% and 81.20%. After tempering the bulk lipid (mimicking the end process of production), the lipid showed amorphous characteristics, with a melting point of ca. 30 °C. The toxicity of SLNs was evaluated in two distinct cell lines (HEPG-2 and Caco-2), showing to be dependent on the concentration of particles in HEPG-2 cells, while no toxicity in was reported in Caco-2 cells. SLNs were stable for 24 h in in vitro human serum albumin (HSA) solution. The resulting SLNs fabricated by double emulsion may provide a promising approach for administration of protein therapeutics and antigens.

The expression of efflux and uptake transporters are regulated by statins in Caco-2 and HepG2 cells

Aim: Statin disposition and response are greatly determined by the activities of drug metabolizing enzymes and efflux/uptake transporters. there is little information on the regulation of these proteins in human cells after statin therapy. In this study, the effects of atorvastatin and simvastatin on mRNA expression of efflux (ABCB1, ABCG2 and ABCC2) and uptake (SLCO1B1, SLCO2B1 and SLC22A1) drug transporters in Caco-2 and HepG2 cells were investigated. Methods: Quantitative real-time PCR was used to measure mRNA levels after exposure of HepG2 and Caco-2 cells to statins. Results: Differences in mRnA basal levels of the transporters were as follows: ABCC2>ABCG2>ABCB1>SLCO1B1>>>SLC22A1>SLC O2B1 for HepG2 cells, and SLCO2B1>>ABCC2>ABCB1>ABCG2>>>SLC22A1 for Caco-2 cells. While for HepG2 cells, ABCC2, ABCG2 and SLCO2B1 mRnA levels were significantly up-regulated at 1, 10 and 20 mu mol/L after 12 or 24 h treatment, in Caco-2 cells, only the efflux transporter ABCB1 was significantly down-regulated by two-fold following a 12 h treatment with atorvastatin. Interestingly, whereas treatment with simvastatin had no effect on mRNA levels of the transporters in HepG2 cells, in Caco-2 cells the statin significantly down-regulated ABCB1, ABCC2, SLC22A1, and SLCO2B1 mRnA levels after 12 or 24 h treatment. Conclusion: These findings reveal that statins exhibits differential effects on mRNA expression of drug transporters, and this effect depends on the cell type. Furthermore, alterations in the expression levels of drug transporters in the liver and/or intestine may contribute to the variability in oral disposition of statins.

Comparison of Bidirectional Lamivudine and Zidovudine Transport Using MDCK, MDCK-MDR1, and Caco-2 Cell Monolayers

Bidirectional transport studies were conducted using Caco-2, MDCK, and MDCK-MDR1 to determine P-gp influences in lamivudine and zidovudine permeability and evaluate if zidovudine permeability changes with the increase of zidovudine concentration and/or by association of lamivudine. Transport of lamivudine and zidovudine separated and coadministrated across monolayers based on these cells were quantified using LC-MS-MS. Drug efflux by P-gp was inhibited using GG918. Bidirectional transport of lamivudine and zidovudine was performed across MDCK-MDR1 and Caco-2 cells. Statistically significant transport decrease in B -> A direction was observed using MDCK-MDR1 for zidovudine and MDCK-MDR1 and Caco-2 for lamivudine. Results show increased transport in B -> A and A -> B directions as concentration increases but data from P(app) increase in both directions for both drugs in Caco-2, decrease in MDCK, and does not change significantly in MDCK-MDR1. Zidovudine transport in A -> B direction increases when coadministrated with increasing lamivudine concentration but does not change significantly in B -> A direction. Zidovudine and lamivudine are P-gp substrates, but results assume that P-gp does not affect significantly lamivudine and zidovudine. Their transport in monolayers based on Caco-2 cells increase proportionally to concentration (in both directions) and zidovudine transport in Caco-2 cell monolayer does not show significant changes with lamivudine increasing concentrations. (C) 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:4413-4419, 2009

Isoflavones in food supplements: chemical profile, label accordance and permeability study in Caco-2 cells

Consumers nowadays are playing an active role in their health-care. A special case is the increasing number of women, who are reluctant to use exogenous hormone therapy for the treatment of menopausal symptoms and are looking for complementary therapies. However, food supplements are not clearly regulated in Europe. The EFSA has only recently begun to address the issues of botanical safety and purity regulation, leading to a variability of content, standardization, dosage, and purity of available products. In this study, isoflavones (puerarin, daidzin, genistin, daidzein, glycitein, genistein, formononetin, prunetin, and biochanin A) from food supplements (n = 15) for menopausal symptoms relief are evaluated and compared with the labelled information. Only four supplements complied with the recommendations made by the EC on the tolerable thresholds. The intestinal bioavailability of these compounds was investigated using Caco-2 cells. The apparent permeability coefficients of the selected isoflavonoids across the Caco-2 cells were affected by the isoflavone concentration and product matrix.

HYPOXIC STRESS, HEPATOCYTES AND CACO-2 VIABILITY AND SUSCEPTIBILITY TO Shigella flexneri INVASION

SUMMARY Inflammation due to Shigella flexneri can cause damage to the colonic mucosa and cell death by necrosis and apoptosis. This bacteria can reach the bloodstream in this way, and the liver through portal veins. Hypoxia is a condition present in many human diseases, and it may induce bacterial translocation from intestinal lumen. We studied the ability of S. flexneri to invade rat hepatocytes and Caco-2 cells both in normoxic and hypoxic microenvironments, as well as morphological and physiological alterations in these cells after infection under hypoxia. We used the primary culture of rat hepatocytes as a model of study. We analyzed the following parameters in normoxic and hypoxic conditions: morphology, cell viability, bacterial recovery and lactate dehydrogenase (LDH) released. The results showed that there were fewer bacteria within the Caco-2 cells than in hepatocytes in normoxic and hypoxic conditions. We observed that the higher the multiplicity of infection (MOI) the greater the bacterial recovery in hepatocytes. The hypoxic condition decreased the bacterial recovery in hepatocytes. The cytotoxicity evaluated by LDH released by cells was significantly higher in cells submitted to hypoxia than normoxia. Caco-2 cells in normoxia released 63% more LDH than hepatocytes. LDH increased 164% when hepatocytes were submitted to hypoxia and just 21% when Caco-2 cells were in the same condition. The apoptosis evaluated by Tunel was significantly higher in cells submitted to hypoxia than normoxia. When comparing hypoxic cells, we obtained more apoptotic hepatocytes than apoptotic Caco-2 cells. Concluding our results contribute to a better knowledge of interactions between studied cells and Shigella flexneri. These data may be useful in the future to define strategies to combat this virulent pathogen.

Estudos sôbre óleos do fígado de cacão Desmobrânquios da família Sphyrnidae, Mir. Rib., gêneros Carcharias, Raf., Galeocerdo, Ranz., e Odontaspis, Shau

1. The author suggests a tecnique for the determination of vitamin A on shark liver oils in industrial plants. The advantages of using oly four ml. of reagent and of permitting a quickly rigorous reading by photoeletric cell, contribute to the possibility of the examination of a great number of samples daily; 2. It is described a survey on the vitamin A content of oils from shark livers, which has been made at the Finishing School Darcy Vargas, Marammaia Is., Rio de Janeiro State. The conclusions are the following: a) Male individuals have showed generally tendency for higher vitamin A pontency oils; b) The size of the fish does not interfere in the vitamin content of the oil (graphic 4); c) The data collected upon 3.085 individuals led to the conclusion that some species are richer in the reservated vitamin although it was possible to catch in the same specie fishes with widely variable potency in vitamin A. One fish belonging to the specie C. lamia produced the highest vitamin potency oil with 167.712 international units per gram; d) The fishing season appears to have no influence on the oils; e) The adventitous food seems to be the most important factor affecting the content of vitamin A of the shark-liver oils; 3. The presence and the quantity of vitamin D in those oils was investigated and two of the determinations are presented.

Conditioned polarized Caco-2 cell monolayers allow to discriminate for the ability of gut-derived microorganisms to modulate permeability and antigen-induced basophil degranulation.

BACKGROUND: Food allergy is a common allergic disorder--especially in early childhood. The avoidance of the allergenic food is the only available method to prevent further reactions in sensitized patients. A better understanding of the immunologic mechanisms involved in this reaction would help to develop therapeutic approaches applicable to the prevention of food allergy. OBJECTIVE: To establish a multi-cell in vitro model of sensitized intestinal epithelium that mimics the intestinal epithelial barrier to study the capacity of probiotic microorganisms to modulate permeability, translocation and immunoreactivity of ovalbumin (OVA) used as a model antigen. METHODS: Polarized Caco-2 cell monolayers were conditioned by basolateral basophils and used to examine apical to basolateral transport of OVA by ELISA. Activation of basophils with translocated OVA was measured by beta-hexosaminidase release assay. This experimental setting was used to assess how microorganisms added apically affected these parameters. Basolateral secretion of cytokine/chemokines by polarized Caco-2 cell monolayers was analysed by ELISA. RESULTS: Basophils loaded with OVA-specific IgE responded to OVA in a dose-dependent manner. OVA transported across polarized Caco-2 cell monolayers was found to trigger basolateral basophil activation. Microorganisms including lactobacilli and Escherichia coli increased transepithelial electrical resistance while promoting OVA passage capable to trigger basophil activation. Non-inflammatory levels of IL-8 and thymic stromal lymphopoietin were produced basolaterally by Caco-2 cells exposed to microorganisms. CONCLUSION: The complex model designed in here is adequate to learn about the consequence of the interaction between microorganisms and epithelial cells vis-a-vis the barrier function and antigen translocation, two parameters essential to mucosal homeostasis. It can further serve as a direct tool to search for microorganisms with anti-allergic and anti-inflammatory properties.

Messenger RNA expression of transporter and ion channel genes in undifferentiated and differentiated Caco-2 cells compared to human intestines.

PURPOSE: The purpose of this work was to study the influence of cell differentiation on the mRNA expression of transporters and channels in Caco-2 cells and to assess Caco-2 cells as a model for carrier-mediated drug transport in the intestines. METHOD: Gene mRNA expression was measured using a custom-designed microarray chip with 750 deoxyoligonucleotide probes (70mers). Each oligomer was printed four times on poly-lysine-coated glass slides. Expression profiles were expressed as ratio values between fluorescence intensities of Cy3 and Cy5 dye-labeled cDNA derived from poly(A) + RNA samples of Caco-2 cells and total RNA of human intestines. RESULTS: Significant differences in the mRNA expression profile of transporters and channels were observed upon differentiation of Caco-2 cells from 5 days to 2 weeks in culture, including changes for MAT8, S-protein, and Nramp2. Comparing Caco-2 cells of different passage number revealed few changes in mRNAs except for GLUT3, which was down-regulated 2.4-fold within 13 passage numbers. Caco-2 cells had a similar expression profile when either cultured in flasks or on filters but differed more strongly from human small and large intestine, regardless of the differentiation state of Caco-2 cells. Expression of several genes highly transcribed in small or large intestines differed fourfold or more in Caco-2 cells. CONCLUSIONS: Although Caco-2 cells have proven a suitable model for studying carrier-mediated transport in human intestines, the expression of specific transporter and ion channel genes may differ substantially.

Evaluation of the interactions between multiwalled carbon nanotubes and caco-2 cells

The aim of this study was to determine whether multiwalled carbon nanotubes (MWNCT) are taken up by and are toxic to human intestinal enterocytes using the Caco-2 cell model. Caco-2 cells were exposed to 50 ?g/ml MWCNT (oxidized or pristine) for 24 h, and experiments were repeated in the presence of 2.5 mg/L natural organic matter. Cells displayed many of the properties that characterize enterocytes, such as apical microvilli, basolateral basement membrane, and glycogen. The cell monolayers also displayed tight junctions and electrical resistance. Exposure to pristine and oxidized MWCNT, with or without natural organic matter, did not markedly affect viability, which was assessed by measuring activity of released lactate dehydrogenase (LDH) and staining with propidium iodide. Ultrastructural analysis revealed some damage to microvilli colocalized with the MWCNT; however, neither type of MWCNT was taken up by Caco-2 cells. In contrast, pristine and oxidized MWCNT were taken up by the macrophage RAW 264.7 line. Our study suggests that intestinal enterocytes cells do not take up MWCNT. [Authors]

A link between FXYD3 (Mat-8)-mediated Na,K-ATPase regulation and differentiation of Caco-2 intestinal epithelial cells.

FXYD3 (Mat-8) proteins are regulators of Na,K-ATPase. In normal tissue, FXYD3 is mainly expressed in stomach and colon, but it is also overexpressed in cancer cells, suggesting a role in tumorogenesis. We show that FXYD3 silencing has no effect on cell proliferation but promotes cell apoptosis and prevents cell differentiation of human colon adenocarcinoma cells (Caco-2), which is reflected by a reduction in alkaline phosphatase and villin expression, a change in several other differentiation markers, and a decrease in transepithelial resistance. Inhibition of cell differentiation in FXYD3-deficient cells is accompanied by an increase in the apparent Na+ and K+ affinities of Na,K-ATPase, reflecting the absence of Na,K-pump regulation by FXYD3. In addition, we observe a decrease in the maximal Na,K-ATPase activity due to a decrease in its turnover number, which correlates with a change in Na,K-ATPase isozyme expression that is characteristic of cancer cells. Overall, our results suggest an important role of FXYD3 in cell differentiation of Caco-2 cells. One possibility is that FXYD3 silencing prevents proper regulation of Na,K-ATPase, which leads to perturbation of cellular Na+ and K+ homeostasis and changes in the expression of Na,K-ATPase isozymes, whose functional properties are incompatible with Caco-2 cell differentiation.

Potentiation of polarized intestinal Caco-2 cell responsiveness to probiotics complexed with secretory IgA.

The precise mechanisms underlying the interaction between intestinal bacteria and the host epithelium lead to multiple consequences that remain poorly understood at the molecular level. Deciphering such events can provide valuable information as to the mode of action of commensal and probiotic microorganisms in the gastrointestinal environment. Potential roles of such microorganisms along the privileged target represented by the mucosal immune system include maturation prior, during and after weaning, and the reduction of inflammatory reactions in pathogenic conditions. Using human intestinal epithelial Caco-2 cell grown as polarized monolayers, we found that association of a Lactobacillus or a Bifidobacterium with nonspecific secretory IgA (SIgA) enhanced probiotic adhesion by a factor of 3.4-fold or more. Bacteria alone or in complex with SIgA reinforced transepithelial electrical resistance, a phenomenon coupled with increased phosphorylation of tight junction proteins zonula occludens-1 and occludin. In contrast, association with SIgA resulted in both enhanced level of nuclear translocation of NF-κB and production of epithelial polymeric Ig receptor as compared with bacteria alone. Moreover, thymic stromal lymphopoietin production was increased upon exposure to bacteria and further enhanced with SIgA-based complexes, whereas the level of pro-inflammatory epithelial cell mediators remained unaffected. Interestingly, SIgA-mediated potentiation of the Caco-2 cell responsiveness to the two probiotics tested involved Fab-independent interaction with the bacteria. These findings add to the multiple functions of SIgA and underscore a novel role of the antibody in interaction with intestinal bacteria.

Role of P-glycoprotein in the uptake/efflux transport of oral vitamin K antagonists and rivaroxaban through the Caco-2 cell model.

Vitamin K antagonists (VKAs) are prescribed worldwide and remain the oral anticoagulant of choice. These drugs are characterized by a narrow therapeutic index and a large inter- and intra-individual variability. P-glycoprotein could contribute to this variability. The aim of this study was to investigate the involvement of P-gp in the transport of acenocoumarol, phenprocoumon and warfarin using an in vitro Caco-2 cell monolayer model. These results were compared with those obtained with rivaroxaban, a new oral anticoagulant known to be a P-gp substrate. The transport of these four drugs was assessed at pH conditions 6.8/7.4 in the presence or absence of the P-gp inhibitor cyclosporine A (10 μM) and the more potent and specific P-gp inhibitor valspodar (5 μM). Analytical quantification was performed by LC/MS. With an efflux ratio of 1.7 and a significant decrease in the efflux (Papp B-A), in the presence of P-gp inhibitors at a concentration of 50 μM, acenocoumarol can be considered as a weak P-gp substrate. Concerning phenprocoumon, the results suggest that this molecule is a poor P-gp substrate. The P-gp inhibitors did not affect significantly the transport of warfarin. The efflux of rivaroxaban was strongly inhibited by the two P-gp inhibitors. In conclusion, none of the three VKAs tested are strong P-gp substrates. However, acenocoumarol can be considered as a weak P-gp substrate and phenprocoumon as a poor P-gp substrate.